Background: Transcutaneous in vivo electroporation is expected to be an effective gene-transfer method for promoting bone regeneration using the BMP-2 plasmid vector. To promote enhanced osteoinduction using this method, we simultaneously transferred cDNAs for BMP-2 and BMP-7, as inserts in the non-viral vector pCAGGS.
Methods: First, an in vitro study was carried out to confirm the expression of BMP-2 and BMP-7 following the double-gene transfer. Next, the individual BMP-2 and BMP-7 plasmids or both
together were injected into rat calf muscles, and transcutaneous electroporation was applied 8 times at 100 V, 50 msec.
Results: In the culture system, the simultaneous transfer of the BMP-2 and BMP-7 genes led to a much higher ALP activity in C2C12 cells than did the transfer of either gene alone. In vivo, ten days after the treatment, soft X-ray analysis showed that muscles that received both pCAGGS-BMP-2 and pCAGGS-BMP-7 had better-defined opacities than those receiving a single gene. Histological examination showed advanced ossification in calf muscles that received the double-gene transfer.
BMP-4 mRNA was also expressed, and RT-PCR showed that its level increased for 3 days in a timedependent manner in the double-gene transfer group. Immunohistochemistry confirmed that BMP-
4-expressing cells resided in the matrix between muscle fibers.
Conclusion: The simultaneous transfer of BMP-2 and BMP-7 genes using in vivo electroporation induces more rapid bone formation than the transfer of either gene alone, and the increased
expression of endogenous BMP-4 suggests that the rapid ossification is related to the induction of
|備考||Digital Object Identifer:10.1186/1471-2474-7-62|
Published with permission from the copyright holder. This is the institute's copy, as published in BMC Musculoskeletal Disorders, 3 August 2006, 7:62.
© 2006 Kawai et al; licensee BioMed Central Ltd.
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